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Articles by Toshiyuki Matsui
Total Records ( 14 ) for Toshiyuki Matsui
  Toshiyuki Matsui , Kyosuke Yokozeki and Hiroyuki Inoue
  The present study was carried out to clone and sequence 1-aminocyclopropane-1-carboxylate synthase (ACC synthase, pBA-ACS) from a sliced moso bamboo shoot using reverse transcription and polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends-PCR (RACE-PCR). The cDNA clone of pBA-ACS was 1793 bp in length and contained a 5'-untranslated region of 126 bp, an open reading frame of 1446 bp encoding 482 amino acids and 3'-untranslated region of 221bp containing stop codone. The pBA-ACS was highly homologous to ACC synthase genes from rice, followed by apple and arabidopsis. In northern analysis, expression of pBA-ACS mRNA was enhanced in wounding tissue until 24 h and coincided with the peak of ACS synthase activity but disagreed with ethylene production. The ACC synthase activity suggested that the increase might be the response to the wounding associated with harvest.
  Pankaj Kumar Bhowmik , Toshiyuki Matsui , Haruo Suzuki and Yusuke Kosugi
  The plant actin cytoskeleton is central to many different sub-cellular processes. Higher plants contain families of actin-encoding genes which are divergent and differentially expressed. Progress in understanding the functions and evolution of plant actins has been hindered by the large size of actin gene families. A cDNA sequence encoding asparagus actin gene was PCR-amplified, cloned and sequenced. Phylogenetic analysis of the actin along with those previously published in the database, indicates that monocots actin genes are monophyletic and underwent duplications during evolution.
  Pankaj Kumar Bhowmik and Toshiyuki Matsui
  Changes in soluble sugar content and the activity of acid invertase (AI), sucrose synthase (SS) and sucrose phosphate synthase (SPS) in the roots of two cultivars of asparagus (Asparagus officinalis L. cvs. Welcome and E414) grown using the mother stalk culture were studied. The primary objective of this study was to understand the effect of long-term harvesting (March-October) on the carbohydrate status of asparagus roots. All the three soluble sugars sucrose, glucose and fructose declined from March to a minimum in May as the new shoots developed. After that root sugar level remained almost unchanged until August and thereafter started to decline again. The activities of both the soluble and cell wall-bound acid invertase increased until May and after that they started to decline. On the other hand, SS and SPS activity decreased to a maximum in May and changed little after that. The activity of acid invertase was higher in ‘E414’ than ‘Welcome’ (WC), whereas those of SS and SPS were higher in ‘WC’ than in ‘E414’, although there were no significant differences in soluble sugar content in the two cultivars.
  Toshiyuki Matsui , Pankaj Kumar Bhowmik and Kyosuke Yokozeki
  Previously isolated cDNA clones encoding glutamine synthetase (GS, EC 6.3.1.2) genes from asparagus and broccoli were observed that the level of transcripts decreased in both plant species during storage. In order to identify factors regulating the postharvest deterioration of another important Asian vegetable bamboo (Phyllostachys edulis) shoot, the present study was conducted to isolate glutamine synthetase (pBA-GS) cDNA clone from a cDNA library prepared from harvested bamboo shoot using reverse transcriptase-polymerase chain reaction. The partial cDNA clone encodes an mRNA of 832 bp and the derived amino acid sequence is highly homologous to GS from rice, barely and maize. Northern blot analysis showed decrease level of pBA-GS mRNA until 9 d at 20°C, which coincided well with enzyme activity and ammonia accumulation. Results demonstrated that GS activity decrease during the postharvest storage of bamboo shoot which is a highly perishable vegetable like asparagus spears and broccoli head and is likely to be a critical feature of their postharvest physiology.
  Toshiyuki Matsui , Pankaj Kumar Bhowmik and Kyosuke Yokozeki
  In plants, phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) catalyzes the conversion of L-phenylalanine to trans cinnamic acid in the first step of phenylpropanoid pathway. We previously isolated cDNA clones encoding PAL from asparagus spears and observed that the level of transcripts correlated well with increase enzyme activity and lignification. In order to identify factors regulating the postharvest deterioration of another important Asian vegetable bamboo (Phyllostachys edulis) shoot, we have now isolated PAL (pBA-PAL), cDNA clone from a cDNA library prepared from harvested bamboo shoot using reverse transcriptase-polymerase chain reaction. The partial cDNA clone encodes an mRNA of 527 bp and the derived amino acid sequence is highly homologous to PAL from rice, barely, maize and asparagus. Northern blot analysis showed increasing trend in the level of pBA-PAL mRNA reaching a peak after 3 d of storage at 20°C, which coincided well with enzyme activity. Our results demonstrate that during the postharvest storage of bamboo shoot PAL activity is regulated by increase level of pBA-PAL mRNA suggesting that the increase might be a response to the wounding associated with harvest.
  Pankaj Kumar Bhowmik and Toshiyuki Matsui
  Not Available
  Toshiyuki Matsui , Pankaj Kumar Bhowmik and Kyosuke Yokozeki
  Actin is one of the most ubiquitous and conserved eukaryotic protein. All eukaryotic cells, from yeast to plants to animals, have an internal framework called the cytoskeleton. The plant actin cytoskeleton is central to many different sub-cellular processes and actin genes have been the focus of numerous scientific studies because of their involvement in basic cellular processes. In order to explain the structure and evolution of monocot actin gene family we have cloned and sequenced a 515 bp cDNA sequence from moso bamboo (Phyllostachys edulis) shoot. Phylogenetic analysis of the bamboo actin along with asparagus actin and those previously published in the database, indicates that monocots actin genes are highly conserved and actin multigene family underwent many gene duplication events early in the evolution of angiosperms.
  Dulal Chandra , Toshiyuki Matsui , Haruo Suzuki , Yusuke Kosugi and Koichi Fujimura
  In order to understand the factors contributing to postharvest deterioration of lettuce, the changes in ammonia content as well as activity and gene expression of GS were investigated in the outer and inner leaves of lettuce head during storage at 20°C. About two times higher ammonia content out of its initial content was found in the outer leaf portion after the end of 96 h storage period. GS activity in this portion declined to about 27% of the initial level by 24 h of storage, while activity in the inner leaf portion almost unchanged throughout the storage. To understand clearly these biochemical changes, a cDNA encoding GS was isolated, cloned and sequenced from lettuce leaves. The partial cDNA clone referred to as LsGS (Lactuca sativa Glutamine Synthetase; AB440673) consisted of 799 nucleotides which showed more than 80% similarity for both nucleotide and amino acid level with the GS genes of other dicotyledonous plant. Northern blot analysis showed that the level of transcript of GS decreased in the outer-leaf portion after 24 h of storage which well correlates with the enzyme activity of this portion. Although, we found no specific trend in GS activity in the inner-leaf portion, the transcript level gradually increased until the end of storage period. The inconsistency between enzyme activity and gene expression may suggest that GS expression in lettuce is controlled by multiple levels of regulations in a tissue-specific manner.
  Dulal Chandra , Toshiyuki Matsui , Haruo Suzuki and Yusuke Kosugi
  To investigate the effects of low temperature on the physiological and biochemical processes, we held two crisphead lettuce (Lactuca sativa L.) cultivars ‘Shizuka` and ‘Cisco` at 5°C for 10 days. Minimum weight loss and lower respiration rate along with a higher hue angle value were recorded from both cultivars. However, cultivar ‘Cisco` showed higher relative weight loss, respiration rate and accumulation of ammonia compared to that of ‘Shizuka`. Except in the leaf portion of ‘Shizuka`, glutamine synthetase (GS, 6.3.1.2) activity did not change significantly. The accumulation of ammonia was supposed to be below the critical level to repress GS activity. However, the highest amount of ammonia content was found at the end of the storage period in all cases. Amination activity of glutamate dehydrogenase (GDH, 1.4.1.2) decreased by day 4 and slightly increased thereafter while the deamination activity did not show any significant change. Overall, GDH-amination activity was remarkably higher than GDH-deamination activity and the leaf portion showed higher GDH-amination activity while the midrib portion showed higher GDH-deamination activity. Between the two cultivars, ‘Cisco` could have a shorter shelf life than ‘Shizuka`. The results suggested that although the visual quality of lettuce was maintained at low temperature, some physio-biochemical changes occurred in this period.
  Toshiyuki Matsui , Takao Lkeuchi and Kazuhide Kawada
  Changes in acid invertase (EC3.2.1.26), fructanase (EC3.2.1.7) activities and sugar distribution were investigated in the different portions of asparagus (Asparagus officinalis. L. cv. E414, HC, WC and S-235, grown in greenhouse) spears harvested in "Spring harvest (2nd and 3rd of March)", "Middle harvest (27th and 29th of May)" and "Summer harvest (27th of August)". The weight of "Middle harvest" asparagus was apt to be higher than "Spring harvest" and "Summer harvest". In soluble and cell wall-bound fractions, the acid invertase and fructanase activities of the two spears increased in the "Middle harvest" concurrent with the most rapid increased in reducing sugar content but with a lower sucrose and fructane contents. In the top portion of ‘HC’, substrate of sucrose was negatively correlated with acid invertase activity in soluble fraction and accounted well for the relation between the substrate and the activity. Due to the existence of fructane in the spear, fructose content was considered to be higher than glucose content. The best harvest time was in the "Middle harvest" since the total sugar content and fresh weight were adequate.
  Pankaj Kumar Bhowmik , Toshiyuki Matsui and Kazuhide Kawada
  We held Asparagus (Asparagus officinalis L.) spears at 1°C for up to 7 and 14 days and then transferred to 25°C to examine the textural and compositional changes that occurred during low temperature storage and subsequent senescence. The preliminary objective of the study was to determine the influence of storage time on the shelf life and edible quality of Asparagus spears when they are transferred to a little bit high temperature to simulate retail display in the super market. A general increase in the amount of fiber of the spears along with the increase of storage time was observed. The toughness determined as the value of breaking force was higher in the bottom portion than in the top portion. The fiber content of the spears increased more when they were transferred to 25°C and the increase was maximum in the spears stored for 14 days at 1°C. There was a gradual loss in sugar content throughout the storage period and the trend of declines were more or less similar for sucrose, glucose and fructose although sucrose was found only in very low concentrations both in the top and bottom portions. The sugar loss was higher in the spears stored for 14 days. Organic acid content also decreased with the increase of storage time. Moreover, the spears stored for 7 days at 1°C remained edible up to 4 days after transferring to 25°C but those stored for 14 days remained only for 2 days which suggested to avoid long time low temperature storage of Asparagus before exporting or simulating retail sale.
  Toshiyuki Matsui , Yousuke Hatase and Kengo Ohobayashi
  We cloned and sequenced 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase, pBA-ACO) from a wounding related moso bamboo (Phyllostachys edulis) shoot using reverse transcription and polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends-PCR (RACE-PCR). The pBA-ACO was highly homologus ACC oxidase genes from submerged deepwater rice, followed by rice and banana, all of which were the wound-inducing genes in monocotyledon plants. In Northern analysis, expression of pBA-ACO mRNA was enhanced in wounding tissue until 16 hr and coincided with the peak of ACC oxidase activity.
  Pankaj Kumar Bhowmik , Toshiyuki Matsui , Fabio Gimena Enriquez , A.K.M. Shameem Alam and Kazuhide Kawada
  We held asparagus (Asparagus officinalis L.) spears at 25°C for up to 5 days after harvest and examined changes in soluble carbohydrates and the activities of enzymes concerned with carbohydrate breakdown in both top and bottom portions of the spears. The acid invertase in soluble fraction showed a higher activity than that in cell wall bound fraction and the top portion of the spear showed a significantly higher soluble acid invertase activity than the bottom portion. But the activity of cell wall bound acid invetase was higher in bottom portion. In both the top and bottom portions soluble acid invertase activity increased during first day of storage and after that it started to decrease gradually up to five days. In case of cell wall bound acid invertase the activity increased during first two days and after that it also started to decrease. Sucrose synthase activity, found higher in bottom portion than top, started to decline from the first day of storage and continued up to five days. But the activity declined rapidly on third day. In case of sucrose phosphate synthase there was no specific inclining or declining pattern of activity in any portion of the spear. Among the soluble sugars fructose and glucose were predomonant and fructose content was significantly higher than the glucose and sucrose. All the three sugars started to decrease from the first day and continued up to five days. Sucrose content was negatively correlated with invertase and sucrose synthase in both portions and accounted well for the relation between the substrate and enzyme activity. But sucrose phohphate synthase activity remained almost constant during the storage period and there was no significant correlation between sugar content and sucrose phosphate synthase activity.
  Toshiyuki Matsui , Yusuke Kosugi , Tomohiro Yanagi , Haruo Suzuki , Pankaj Kumar Bhowmik and Sutevee Sukarakarn
  The genetic relationship of 13 oriental melon (Cucumis melo L.) was determined using random amplified polymorphic DNA (RAPD) analysis with 12 dodecamer oligonucleotide primers. Representative cultivars could be divided into two major groups at 0.756 similarity value. One group included 7 makuwa melons namely, ‘ Nara-1-gou ’, ‘ Tiger-melon ’, ‘ Kinpyou ’, ‘ Kiku-melon ’, ‘ Nashi-makuwa ’, ‘ Shouwa-melon ’ and 4 conomon melons namely, ‘ Numame-shirouri ’, ‘ Katsura-ohoshirouri ’, ‘ Tokyo-ohoshirouri ’, ‘ Kuromonao-ohoshirouri ’. The other major group included 2 makuwa cultivars namely ‘ Ginsen ’ and ‘ Wasegin ’. The RAPD showed that conomon melon belonged to same species as makuwa melon. It seemed that, makuwa melon is an allogamous plant which hybridizes easily with table-melons and conomon melon and it might have hybridized with conomon melon in the distant past.
 
 
 
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